替莫唑胺对U251干细胞细胞周期及细胞增殖的影响

The effect of temozolomide on cell cycle and proliferation of cancer stem cells isolated from glioblastoma U251cells

目的研究化疗药物TMZ对胶质瘤U251细胞和U251干细胞的细胞周期以及细胞增殖活性的影响。方法从胶质瘤U251细胞系中用免疫磁珠法分选出胶质瘤干细胞,CCK-8检测细胞增殖速度。用不同浓度的化疗药物TMZ分别干预U251及其干细胞,分别在干预后24h、48h、72h收集细胞,流式细胞仪检测U251及其干细胞的细胞周期变化,CCK-8检测TMZ干预前后细胞增殖变化。结果 U251细胞增殖速度明显高于U251干细胞。TMZ能使U251细胞的细胞周期停留在G2/M期,U251干细胞则停留在S期,细胞增殖停止。随着药物浓度增高和干预时间的延长,U251细胞及干细胞细胞周期停滞,抑制细胞增殖的效果越明显。相同作用时间内同一药物浓度,TMZ对干细胞的增殖抑制作用弱于U251细胞。结论 TMZ能有效抑制U251及其干细胞,使细胞周期停滞,降低胶质瘤细胞及其干细胞的增殖速度,抑制肿瘤细胞增殖。TMZ对抑制U251细胞增殖的作用强于U251干细胞,这可能是肿瘤耐药的机制不同。

Objective To analyse the change of cell cycle and proliferation in the glioblastoma U251 cells and it ＇s cancer stem cells with chemotherapy drug temozolomide intervention.Methods The cancer stem cells from glioblastoma U251 cells were isolated by using immune magnetic beads.The change of cell cycle and cell proliferation rate was detected by flow cytometric and CCK-8 after the intervention with different drug concentrations（0 μmol/L,25μmol/L,50μmol/L,100μmol/L,200μmol/L,400μmol/L）at different time points（24h,48h,72h）.Results The CD133＋stem cells speed of cell proliferation was significantly lower than U251 cells in the same time and condition of culture.The cell cycle of U251 cells stayed in the G2/M period and it＇s cancer stem cells stayed in S period respectively after drug temozolomide intervention.The cell proliferation rate was obviously inhibited with the increasing of drug concentration and intervention time.But the cell proliferation inhibition of stem cells was weaker than U251 cells under the same drug concentration and the same intervention time.Conclusion The cell cycle and cell proliferation speed were inhibited after intervention with temozolomide,but the inhibition was stronger in U251 cells than stem cells after TMZ intervention with the same condition,which may be the different mechanism of drug resistance between U251 cells and stem cells.